Equine Embryonic Stem Cell-Derived Tenocytes are Insensitive to a Combination of Inflammatory Cytokines and Have Distinct Molecular Responses Compared to Primary Tenocytes

Smith, Emily J; Beaumont, Ross E; Dudhia, Jayesh; Guest, Deborah J

Abstract

Tissue fibrosis following tendon injury is a major clinical problem due to the increased risk of re-injury and limited treatment options; however, its mechanism remains unclear. Evidence suggests that insufficient resolution of inflammation contributes to fibrotic healing by disrupting tenocyte activity, with the NF-κB pathway being identified as a potential mediator. Equine embryonic stem cell (ESC) derived tenocytes may offer a potential cell-based therapy to improve tendon regeneration, but how they respond to an inflammatory environment is largely unknown. Our findings reveal for the first time that, unlike adult tenocytes, ESC-tenocytes are unaffected by IFN-γ, TNFα, and IL-1β stimulation; producing minimal changes to tendon-associated gene expression and generating 3-D collagen gel constructs indistinguishable from unstimulated controls. Inflammatory pathway analysis found these inflammatory cytokines failed to activate NF-κB in the ESC-tenocytes. However, NF-κB could be activated to induce changes in gene expression following stimulation with NF-κB pharmaceutical activators. Transcriptomic analysis revealed differences between cytokine and NF-κB signalling components between adult and ESC-tenocytes, which may contribute to the mechanism by which ESC-tenocytes escape inflammatory stimuli. Further investigation of these molecular mechanisms will help guide novel therapies to reduce fibrosis and encourage superior tendon healing.

Citation

Smith, E. J., Beaumont, R. E., Dudhia, J., & Guest, D. J. (in press). Equine Embryonic Stem Cell-Derived Tenocytes are Insensitive to a Combination of Inflammatory Cytokines and Have Distinct Molecular Responses Compared to Primary Tenocytes. Stem Cell Reviews and Reports, https://doi.org/10.1007/s12015-024-10693-8

• Journal Article Type: Article
• Acceptance Date: Feb 6, 2024
• Online Publication Date: Feb 24, 2024
• Deposit Date: Feb 26, 2024
• Publicly Available Date: Feb 27, 2024
• Journal: Stem Cell Reviews and Reports
• Print ISSN: 1550-8943
• Electronic ISSN: 1558-6804
• Publisher: Springer
• Peer Reviewed: Peer Reviewed
• DOI: https://doi.org/10.1007/s12015-024-10693-8
• Keywords: General Medicine
• Additional Information: Accepted: 6 February 2024; First Online: 24 February 2024; : ; : The authors have no competing interests to declare that are relevant to the content of this article.; : The equine cells used in this study were isolated with approval from the Royal Veterinary College Clinical Research Ethical Review Board (URN 2020 2017–2) and the Animal Health Trust Ethical Review Committee (AHT_02_2012).; : Not applicable.; : Not applicable.; ; Student co-author = Emily Smith - PhD student